LOCUS       Exported                6964 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Intersectional viral expression of Arch3.3-EYFP in cells expressing 
            Cre AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-nEF-Con/Foff 2.0-Arch3.3-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6964)
  AUTHORS   Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M,
            Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K
  TITLE     Comprehensive Dual- and Triple-Feature Intersectional Single-Vector 
            Delivery of Diverse Functional Payloads to Cells of Behaving 
            Mammals.
  JOURNAL   Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003.
  PUBMED    32574559
REFERENCE   2  (bases 1 to 6964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6964
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        188..399
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             412..680
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    708..741
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      834..843
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    1136..1169
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACCT). loxN sites are compatible with each 
                     other, but incompatible with loxP or lox2272 sites."
     protein_bind    1220..1253
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     primer_bind     1366..1387
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1430..1486)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(2166..2199)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACCT). loxN sites are compatible with each 
                     other, but incompatible with loxP or lox2272 sites."
     protein_bind    complement(2250..2283)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     primer_bind     complement(2505..2524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2664..2915
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     2852..2873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2949..2982)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3091..3679
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3144..3164)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3562..3573)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3711..4187
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3910..3929)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4227..4367
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4227..4356
                     /label=AAV2 ITR
     rep_origin      4442..4897
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4529..4548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4739..4760
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4914..4933)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5033..5055
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5093..5111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5179..5283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5284..6144
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5502..5521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6315..6903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6804..6823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagga tctgcgatcg ctccggtgcc
      181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      241 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
      301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
      361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
      421 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
      481 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
      541 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
      601 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
      661 gttctgcgcc gttacagatc caagctgtga ccggcgccta cggatccgaa gttcctattc
      721 tctagaaagt ataggaactt cgcagagtag tagctggatt gtagctgcta ttagcaatct
      781 aaaacctctt agaagttcct attcttcaaa aggtatagga acttcggcgc gccgccacca
      841 tggaccccat cgctctgcag gctggttacg acctgctggg tgacggcaga cctgaaactc
      901 tgtggctggg catcggcact ctgctgatgc tgattggaac cttctacttt ctggtccgcg
      961 gatggggagt caccgataag gatgcccggg aatattacgc tgtgactatc ctggtgcccg
     1021 gaatcgcatc cgccgcatat ctgtctatgt tctttggtat cgggcttact gaggtgaccg
     1081 tcgggggcga aatgttggat atctattatg ccaggtaagt gtcggggttg agctcataac
     1141 ttcgtatagt ataccttata cgaagttatt ctttgcacca ttctaaagaa taacagtgat
     1201 aatttctggg ttaaggcaaa taacttcgta taggatactt tatacgaagt tatggtaccc
     1261 ctcctgtact caccttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     1321 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1381 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacg ggaccggggt
     1441 tttcttccac gtctcctgct tgctttaaca gagagaagtt cgtggctccg gagccgggcg
     1501 cgcccacgtt gatgtcgatc tggtccaggg ggatgtactc gccctcgctg gtgatcctgc
     1561 tcttgtcggc ggcactgaca tcggcaccgg cactgggttc tggtgcctcg gtgtcgccca
     1621 gaatagcccg ggatctcaac aggataaagc caaagccgac cttggcagtc acgtccaaca
     1681 ccataaacag cagagtttcg atgcccaggc ccaccacgcc agcgccctca gtgcctatga
     1741 tccacaggat agggtaagcg gtccacagca ccaagaccag agctgtcagg gtgttaaagg
     1801 tagatgccac ctcggggccc cgctcctttg cagcagatcg cagggatgta gccagaaaat
     1861 agagcaccac tatcatgcaa attgtagaga acaaccacca actgtatctg gctatggccg
     1921 tgtggctcaa ggctccgatg aggccagtga cgatcatcag ggcgtccaca cccaccaggg
     1981 tgccgatggt cacccgatcc accttagcga gaagggccag atccagcagc agaagtgggg
     2041 tggtaaacag ccagtcggcg tacctaaaca catgagaaat agagtcataa gcacatgggt
     2101 cacatacagg agattgtata taacattaat acaattttat aaaaaaaagg gggggcacaa
     2161 ctagtataac ttcgtataag gtatactata cgaagttatt aagaggtttc atattgctaa
     2221 tagcagctac aatccagcta ccattctgca taacttcgta taaagtatcc tatacgaagt
     2281 tatgtcgact ggagagtggc cagcccttct catgttcaga gaacatggtt aactggttaa
     2341 gtcatgtcgt cccacagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     2401 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2461 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2521 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2581 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2641 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2701 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2761 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
     2821 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2881 ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac gaggtgtaat
     2941 gagctagcga agttcctata ctttctagag aataggaact tctaagcaga caatcttgta
     3001 atacatcagc agctagtaga ttgctccagc tagaagttcc tatacctttt gaagaatagg
     3061 aacttcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     3121 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3181 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3241 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3301 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3361 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3421 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3481 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3541 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3601 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3661 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
     3721 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     3781 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     3841 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     3901 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     3961 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     4021 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     4081 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     4141 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     4201 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4261 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4321 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4381 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4441 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4501 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4561 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4621 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4681 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4741 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     4801 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4861 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     4921 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4981 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5041 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5101 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5161 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5221 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5281 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5341 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5401 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5461 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5521 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5581 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5641 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5701 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5761 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5821 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5881 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5941 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6001 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6061 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6121 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6181 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6421 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6961 atgt
//
